Tryptic cleavage as a probe of conformational differences between active and inactive forms of ornithine aminotransferase.

Treatment of ornithine aminotransferase with trypsin resulted in rapid and complete loss of enzyme activity in a process that coincided with a reduction in subunit Mr of about 3000. However, electrophoresis showed that a substantial proportion of the enzyme had not been digested. The component of the preparation of ornithine aminotransferase that was insusceptible to trypsin contained a naturally occurring but enzymically inactive form of the enzyme, and when this had been removed, the remaining fully active enzyme was completely digested. Irreversible inactivation with a substrate analogue made all of the enzyme insusceptible to trypsin. The hydrolyzed enzyme still underwent a very slow half-reaction with ornithine. Sequence analysis of the truncated protein, prepared by blotting from electrophoretic gels, showed that hydrolysis had occurred at peptide bond Lys26-Tyr27.